CN108308031A - A kind of pale reddish brown trident bletilla seed tissue cultural method - Google Patents
A kind of pale reddish brown trident bletilla seed tissue cultural method Download PDFInfo
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- CN108308031A CN108308031A CN201810337579.9A CN201810337579A CN108308031A CN 108308031 A CN108308031 A CN 108308031A CN 201810337579 A CN201810337579 A CN 201810337579A CN 108308031 A CN108308031 A CN 108308031A
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/008—Methods for regeneration to complete plants
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/001—Culture apparatus for tissue culture
Abstract
The present invention provides a kind of pale reddish brown trident bletilla seed tissue cultural method, belongs to bletilla field of tissue culture.A kind of pale reddish brown trident bletilla seed tissue cultural method provided by reserve seed for planting, capsule selection, capsule processing, aseptic seeding, seed sprout and from proliferative induction, strong plantlets and rootage, hardening, the bottle outlet sprouted wash seedling, sterilisation step carry out tissue cultivating and seedling.The present invention improves bulb, the leaf blade size of pale reddish brown trident bletilla seed tissue culture seedling, and blade base is more sturdy, improves the plantation survival rate of bletilla.
Description
Technical field
The invention belongs to bletilla field of tissue culture, and in particular to a kind of pale reddish brown trident bletilla seed tissue cultural method.
Background technology
Bletilla is orchid family bletilla category herbaceos perennial, the high 18-60cm of plant；Pseudobulb oblate spheroid, upper mask water chestnut
As annulus, rich viscosity；Stem thickness is strong, rigidness.Bletilla Chang Yesheng is in hills, low mountain stream side and the moon covers thick grass or hayashishita is wet
Ground is distributed in the ground such as provinces and regions and Japan on the south China Qinling Mountains；It is used as medicine with stem tuber, there is tonifying lung hemostasis, detumescence and promoting granulation and other effects,
Also it is widely used in the industries such as medicine, beauty, coating and printing and dyeing simultaneously.In actual production, artificial bletilla cultivation is chronically at " just
Harvest-reserve seed for planting-cultivate again on the spot on the spot in ground " original loop state, bletilla quality degenerates year by year；In exploitation introduces a collection cultivation side
Face, the presence of the high insufficient and invalid seedling phenomenon of the bletilla tissue-cultured seedling death rate using tissue-cultured seedling to alleviating the extensive industry of bletilla
Change plantation and produces certain restriction.Pale reddish brown trident bletilla be a kind of pattern be purple, the Herba Phaii tankervilliaes kind that mode of appearance is trident,
Existing pale reddish brown trident bletilla method for tissue culture turns out the bletilla bottle seedling bulb come and blade is smaller, and late growth is slow,
Survival rate is low.
Invention content
Present invention aim to address above-mentioned deficiencies, provide a kind of pale reddish brown trident bletilla seed tissue cultural method.
To achieve the above object, technical solution of the invention is：A kind of pale reddish brown trident bletilla seed tissue culture side
Method, it is characterised in that carry out as follows：
The first step is reserved seed for planting：The plant of growth selection stalwartness, and without the pale reddish brown trident bletilla striata maternal plant of other types in diameter 1Km,
Florescence, spontaneous pollination select the capsule of mature and plump to harvest when capsule starts to turn yellow.
Second step, capsule selection：Selection mature and plump no disease and pests harm, has no mechanical damage, the capsule without natural cracking.
Third walks, capsule processing：First capsule both ends are handled, with scissors carpopodium length to 1cm, end black
Wrappage gently removes, but to ensure that capsule is complete, not crack；The dust about 1 on surface is cleaned with the small flowing water of Drinking Water again
Hour, the moisture content on surface is dried, is wrapped up with clean-keeping paper；Treated, and capsule is impregnated 30 seconds with 75% alcohol, then uses sterile water
Cleaning 2~3 times, then with the mercuric chloride of 0.1% concentration immersion 30min, then with sterile water wash 4~5 times, after on alcolhol burner flame
Calcination 1S~2S, it is spare.
4th step, aseptic seeding：Aseptically, the capsule carpopodium disinfected is clamped with tweezers, uses scalpel
Capsule top is cut, capsule is placed in the shake in culture bottle gently, one layer of seed of uniform broadcasting is in MS media surfaces, immediately
Seal bottleneck.
5th step, seed are sprouted and from the proliferative inductions sprouted：It is 23 DEG C~27 DEG C that the culture bottle of sowing, which is placed in temperature,
In clean environment, cultivated after a week under the conditions of light irradiation time is 8 hours/day, intensity of illumination is 2000lux~3000lux, then
Under conditions of light irradiation time is 10 hour/day, continues culture 35~45 days and form protocorms and from sprouting.
6th step, strong plantlets and rootage：Culture bottle carries out high temperature sterilization after injecting the proprietary culture mediums of 95~115ml；After sterilization
Selection sprout Multiple Buds that are healthy, pollution-free, growing the high 0.8~1.2cm of two panels leaflet, bud, which are placed on, is marked with proprietary culture medium
It is cultivated in culture bottle, carries out strong plantlets and rootage；The intensity of illumination of culture environment is 2000lux~3000lux, and light irradiation time is 10 small
When/day, it cultivates 120 days~135 days, temperature is 23 DEG C~27 DEG C.
7th step, hardening：Seedling is taken exercise under 15 DEG C~35 DEG C of temperature, illumination 2000lux~20000lux range of condition
After 15 days~20 days, with kind of a seedling leaf, bulb turn it is dark green can bottle outlet.
8th step, bottle outlet wash seedling：Bottleneck is opened, the seedling after hardening is gently pressed from both sides out with tweezers, uses flowing water
Under cleaned, clean root attachment culture medium.
9th step, disinfection：Cleaned seedling broad-spectrum germicide is diluted into spraying or impregnates 5s~10s；At seedling
Reason is placed on shady and cool place and divides storage and transplanted in time.
The special culture media configuration method component by weight：MS culture mediums are 1000 parts, 6BA is 0.5~1.5 part,
NAA is 0.5~1.5 part, powdered carbon is 0.5 part, banana is 50 parts of mixing, and it is 5.8 to add NaOH and adjust pH value.
A kind of pale reddish brown trident bletilla seed tissue cultural method of the offer of the present invention, by long-term experiment and real work
Tissue culture experience improves pale reddish brown trident bletilla seed tissue training by the mating reaction of special culture media each component and each step
Bulb, the leaf blade size of seedling are supported, blade base is more sturdy, improves the plantation survival rate of bletilla.
Specific implementation mode
A kind of pale reddish brown trident bletilla seed tissue cultural method of the offer of the present invention is specifically carried out by following examples.
The first step is reserved seed for planting：The plant of growth selection stalwartness, and it is female without the pale reddish brown trident bletilla striata of other types in diameter 1Km
Strain, at the florescence, spontaneous pollination selects the capsule of mature and plump to harvest when capsule starts to turn yellow.
Second step, capsule selection：Selection mature and plump no disease and pests harm, has no mechanical damage, the capsule without natural cracking.
Third walks, capsule processing：First capsule both ends are handled, with scissors carpopodium length to 1cm, end black
Wrappage gently removes, but to ensure that capsule is complete, not crack；The dust about 1 on surface is cleaned with the small flowing water of Drinking Water again
Hour, the moisture content on surface is dried, is wrapped up with clean-keeping paper；Treated, and capsule is impregnated 30 seconds with 75% alcohol, then uses sterile water
Cleaning 2~3 times, then with the mercuric chloride of 0.1% concentration immersion 30min, then with sterile water wash 4~5 times, after on alcolhol burner flame
Calcination 1S~2S, it is spare.
4th step, aseptic seeding：Aseptically, the capsule carpopodium disinfected is clamped with tweezers, uses scalpel
Capsule top is cut, capsule is placed in the shake in culture bottle gently, one layer of seed of uniform broadcasting is in MS media surfaces, immediately
Seal bottleneck.
5th step, seed are sprouted and from the proliferative inductions sprouted：It is 23 DEG C~27 DEG C that the culture bottle of sowing, which is placed in temperature,
In clean environment, cultivated after a week under the conditions of light irradiation time is 8 hours/day, intensity of illumination is 2000lux~3000lux, then
Under conditions of light irradiation time is 10 hour/day, continues culture 35~45 days and form protocorms and from sprouting.
6th step, strong plantlets and rootage：Culture bottle carries out high temperature sterilization after injecting the proprietary culture mediums of 95~115ml；After sterilization
Selection sprout Multiple Buds that are healthy, pollution-free, growing the high 0.8~1.2cm of two panels leaflet, bud, which are placed on, is marked with proprietary culture medium
It is cultivated in culture bottle, carries out strong plantlets and rootage；The intensity of illumination of culture environment is 2000lux~3000lux, and light irradiation time is 10 small
When/day, it cultivates 120 days~135 days, temperature is 23 DEG C~27 DEG C.
7th step, hardening：Seedling is taken exercise under 15 DEG C~35 DEG C of temperature, illumination 2000lux~20000lux range of condition
After 15 days~20 days, with kind of a seedling leaf, bulb turn it is dark green can bottle outlet.
8th step, bottle outlet wash seedling：Bottleneck is opened, the seedling after hardening is gently pressed from both sides out with tweezers, uses flowing water
Under cleaned, clean root attachment culture medium.
9th step, disinfection：Cleaned seedling broad-spectrum germicide is diluted into spraying or impregnates 5s~10s；At seedling
Reason is placed on shady and cool place and divides storage and transplanted in time.
The special culture media configuration method component by weight：MS culture mediums are 1000 parts, 6BA is 0.5~1.5 part,
NAA is 0.5~1.5 part, powdered carbon is 0.5 part, banana is 50 parts of mixing, and it is 5.8 to add NaOH and adjust pH value.
The culture comparing result of Shucheng County, Anhui Province tissue culture planting base：The ball of traditional pale reddish brown trident bletilla tissue culture seedling
Stem diameter is 5~7mm, and leaf width is 2~5mm, a diameter of 1~1.5mm of blade base, plants survival rate 60~75%；The present invention carries
A diameter of 8~the 14mm of bulb for the seedling that the pale reddish brown trident bletilla tissue culture method supplied is cultivated, leaf width are 5~10mm, blade base
A diameter of 3~the 3.5mm in portion, leaf color is greener, plants survival rate 98%, growing way is obviously superior compared with conventional method.
Above-described embodiment is the technical approach and technical characterstic for illustrating the present invention, in the art common the purpose is to allow
Technical staff cans understand the content of the present invention and implement it accordingly, and it is not intended to limit the scope of the present invention.Every
The equivalent changes or modifications made according to the essence of the content of present invention should all cover within the scope of the present invention.
Claims (2)
1. a kind of pale reddish brown trident bletilla seed tissue cultural method, it is characterised in that：It carries out as follows：
The first step is reserved seed for planting：The plant of growth selection stalwartness, and without the pale reddish brown trident bletilla striata maternal plant of other types in diameter 1Km,
Florescence, spontaneous pollination select the capsule of mature and plump to harvest when capsule starts to turn yellow；
Second step, capsule selection：Selection mature and plump no disease and pests harm, has no mechanical damage, the capsule without natural cracking；
Third walks, capsule processing：First capsule both ends are handled, with scissors carpopodium length to 1cm, the package of end black
Object gently removes, but to ensure that capsule is complete, not crack；The dust about 1 hour on surface is cleaned with the small flowing water of Drinking Water again,
The moisture content for drying surface, is wrapped up with clean-keeping paper；Treated capsule is impregnated 30 seconds with 75% alcohol, then with sterile water wash 2
~3 times, then 30min is impregnated with the mercuric chloride of 0.1% concentration, then with sterile water wash 4~5 times, after on alcolhol burner flame calcination 1S
~2S, it is spare；
4th step, aseptic seeding：Aseptically, the capsule carpopodium disinfected is clamped with tweezers, is cut with scalpel
Capsule is placed in the shake in culture bottle gently by capsule top, and one layer of seed of uniform broadcasting seals immediately in MS media surfaces
Bottleneck；
5th step, seed are sprouted and from the proliferative inductions sprouted：The culture bottle of sowing is placed in the cleaning that temperature is 23 DEG C~27 DEG C
In environment, cultivated after a week under the conditions of light irradiation time is 8 hours/day, intensity of illumination is 2000lux~3000lux, then in light
According to when a length of 10 hour/day under conditions of, continue culture and form within 35~45 days protocorms and from sprouting；
6th step, strong plantlets and rootage：Culture bottle carries out high temperature sterilization after injecting the proprietary culture mediums of 95~115ml；It is selected after sterilization
Sprout Multiple Buds that are healthy, pollution-free, growing the high 0.8~1.2cm of two panels leaflet, bud are placed on the culture for being marked with proprietary culture medium
It is cultivated in bottle, carries out strong plantlets and rootage；The intensity of illumination of culture environment be 2000lux~3000lux, light irradiation time be 10 hours/
It, cultivates 120 days~135 days, and temperature is 23 DEG C~27 DEG C；
7th step, hardening：Seedling is taken exercise 15 days under 15 DEG C~35 DEG C of temperature, illumination 2000lux~20000lux range of condition
After~20 days, with kind of a seedling leaf, bulb turn it is dark green can bottle outlet；
8th step, bottle outlet wash seedling：Open bottleneck, the seedling after hardening is gently pressed from both sides out with tweezers, under flowing water into
Row cleaning, cleans the culture medium of root attachment；
9th step, disinfection：Cleaned seedling broad-spectrum germicide is diluted into spraying or impregnates 5s~10s；After seedling process
Shady and cool place is placed in divide storage and transplanted in time.
2. a kind of pale reddish brown trident bletilla seed tissue cultural method as described in claim 1, it is characterised in that：Described is special
Culture medium configuration method component by weight：MS culture mediums are 1000 parts, 6BA is 0.5~1.5 part, NAA is 0.5~1.5 part, charcoal
Powder is 0.5 part, banana is 50 parts of mixing, and it is 5.8 to add NaOH and adjust pH value.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201810337579.9A CN108308031A (en) | 2018-04-16 | 2018-04-16 | A kind of pale reddish brown trident bletilla seed tissue cultural method |
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| CN201810337579.9A CN108308031A (en) | 2018-04-16 | 2018-04-16 | A kind of pale reddish brown trident bletilla seed tissue cultural method |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109169275A (en) * | 2018-09-13 | 2019-01-11 | 甘肃源宜生物科技有限公司 | A kind of pale reddish brown trident bletilla striata tissue-culturing rapid propagation culture medium and method |
| CN109452171A (en) * | 2018-11-26 | 2019-03-12 | 丽江海贝瑞生物科技有限公司 | A kind of tissue culture method of the sterile induction plant regeneration of pale reddish brown trident bletilla striata seeds |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109169275A (en) * | 2018-09-13 | 2019-01-11 | 甘肃源宜生物科技有限公司 | A kind of pale reddish brown trident bletilla striata tissue-culturing rapid propagation culture medium and method |
| CN109452171A (en) * | 2018-11-26 | 2019-03-12 | 丽江海贝瑞生物科技有限公司 | A kind of tissue culture method of the sterile induction plant regeneration of pale reddish brown trident bletilla striata seeds |
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Application publication date: 20180724 |